US Environmental Protection Agency
Office of Pesticide Programs
Office of Pesticide Programs
Microbiology Laboratory
Environmental Science Center, Ft. Meade, MD
Standard Operating Procedure for
Verification of Digital Pipettes
SOP Number: QC-19-09
Date Revised: 03-05-18
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SOP No. QC-19-09
Date Revised 03-05-18
Page 1 of 8
SOP Number
QC-19-09
Title
Verification of Digital Pipettes
Scope
Describes process for verification of digital pipettes.
Application
Pipettes are calibrated annually by an ISO 17025 accredited vendor
and are verified in-house using the gravimetric procedure as
necessary.
Approval Date
SOP Developer:
Print Name:
SOP Reviewer
Print Name:
Quality Assurance Unit
Print Name:
Branch Chief
Print Name:
Date SOP issued:
Controlled copy
number:
Date SOP withdrawn:
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SOP No. QC-19-09
Date Revised 03-05-18
Page 2 of 8
TABLE OF CONTENTS
Contents Page Number
1.
DEFINITIONS
3
2.
HEALTH AND SAFETY
3
3.
PERSONNEL QUALIFICATIONS AND TRAINING
3
4.
INSTRUMENT CALIBRATION
3
5.
SAMPLE HANDLING AND STORAGE
3
6.
QUALITY CONTROL
3
7.
INTERFERENCES
3
8. NON-CONFORMING DATA
3
9.
DATA MANAGEMENT
3
10.
CAUTIONS
3
11.
SPECIAL APPARATUS AND MATERIALS
3
12.
PROCEDURE AND ANALYSIS
4
13.
DATA ANALYSIS/CALCULATIONS
5
14.
FORMS AND DATA SHEETS
5
15.
REFERENCES
5
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SOP No. QC-19-09
Date Revised 03-05-18
Page 3 of 8
1. Definitions
Abbreviations/definitions are provided in the text.
1. Verification failure = Verification of pipette exceeds ±5% of the target
volume.
2. Ideal volume = Target volume; actual volume being measured, corrected
for temperature using the density of water at 21.0°C (0.997995 g/mL, see
section 15.1).
2. Health and
Safety
Follow procedures specified in SOP MB-01, Laboratory Biosafety. The Study
Director and/or lead analyst should consult the Safety Data Sheet for specific
hazards associated with products.
3. Personnel
Qualifications
and Training
Refer to SOP ADM-04, OPP Microbiology Laboratory Training.
4. Instrument
Calibration
Refer to SOP EQ-03 (weigh balances) for details on method and frequency of
calibration.
5. Sample
Handling and
Storage
None.
6. Quality Control
For quality control purposes, the required information is documented on the
appropriate form(s) (see section 14).
7. Interferences
None.
8. Non-
conforming
Data
1. Manage non-conforming data consistent with SOP ADM-07, Non-
Conformance Reports.
2. Do not use pipettes if the inaccuracy exceeds ±5% of the target volume.
For corrective actions, see section 12.3.
9. Data
Management
1. Archive data consistent with SOP ADM-03, Records and Archives.
2. Maintain an inventory of pipettes electronically using a Microsoft Excel
spreadsheet (refer to section 14). After each addition to or deletion from
the inventory, file a hard copy of the pipette inventory in the Pipette
Verification and Calibration Record Book.
10. Cautions
1. If a pipette is dropped or damaged, successfully verify it in-house using
the gravimetric procedure prior to use.
2. If a pipette fails an in-house verification assessment, do not use it until it
has been repaired (if necessary) and recalibrated by an ISO 17025
accredited vendor.
11. Special
Apparatus and
1. Calibrated balances. Capable of measuring 0.01 g for verifying pipettes
with volumes greater than or equal to 1 mL and 0.001 g for verifying
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SOP No. QC-19-09
Date Revised 03-05-18
Page 4 of 8
Materials
pipettes with volumes less than 1 mL.
2. Pipettes.
a. Rainin Adjustable Volume Pipettes
b. Gilson Microman Positive Displacement Pipettes
c. Gilson Distriman Continuously Adjustable Volume Repetitive
Pipette
d. Eppendorf Repeater Pipette
12. Procedure and
Analysis
12.1 Pipette
Verification
Requirements
a. Annually verify and service pipettes using an ISO 17025 accredited
vendor.
b. If a pipette is dropped or broken, verify the pipette using the
gravimetric verification procedure (see section 10).
c. Record the annual verification results and if necessary, gravimetric
analysis results on the Pipette Verification Record Sheet (refer to
section 14) and file in the Pipette Verification and Calibration Log
Book. The Pipette Verification Record Sheet is based on the
inventory of pipettes and may change over time.
12.2 Gravimetric
Verification
Procedure
a. Record all pertinent information for the gravimetric verification
procedure on the Pipette Verification - Gravimetric Analysis Form
(refer to section 14).
b. In advance of testing, fill a container with de-ionized water and allow
it to equilibrate to room temperature in the same laboratory with the
balance. List the balance that will be used on the appropriate form.
c. Place a small container on the balance and record its mass on the
appropriate form.
d. Using the pipette to be verified, aspirate an aliquot of DI water from
the sample aliquot container and dispense into the small container.
e. Record the mass on the appropriate form.
f. Follow the procedure in 12.2d-e for each subsequent sample
addition; measure at least 5 samples. Do not tare between samples.
After each new sample addition, record the mass on the appropriate
form.
g. Input the measurements and other appropriate information into the
Pipette Verification - Gravimetric Analysis Spreadsheet.
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SOP No. QC-19-09
Date Revised 03-05-18
Page 5 of 8
h. Verify that the percent inaccuracy is within ±5%. If the percent
inaccuracy is outside of this range, remove the pipette from use until
it is repaired and calibrated by a vendor.
„ , . (x-ideal volume)x 100
i. Percent Inaccuracy = -
ideal volume
12.3 Pipette
Verification
Record Sheet
a. From the data on the calibration certificate for each pipette, record
the following on the Pipette Verification Record Sheet:
i. Verification Status: record as "pass" or "fail." Record status
as "pass" if the percent inaccuracy from gravimetric analysis
is within ±5% or if vendor calibration certificate indicates
acceptable results, otherwise record as "fail."
ii. Corrective Action: record as "yes" or "no." If the verification
status is "pass," record as "no." Record as "yes" if the
verification status is "fail," describe the action taken at the
bottom of the page, and notify the Quality Assurance Officer
(QAO). The QAO must determine if there was any impact on
work conducted using the failed pipet.
b. The Pipette Verification Record Sheet may be completed
electronically or by hand.
13. Data Analysis/
Calculations
1. None
14. Forms and Data
Sheets
1. Sample Pipette Verification Record Sheet
2. Sample Pipette Inventory
3. Test Sheets. Test sheets are stored separately from the SOP under the
following file names:
MLB Pipette Inventory QC-19-09 Fl.xlsx
Pipette Verification Record Sheet QC-19-09 F2.xlsx
Pipette Verification - Gravimetric Analysis Form QC-19-09 F3.xlsx
Pipette Verification - Gravimetric Analysis 1
„ r A J QC-19-09 F4.xlsx
Spreadsheet -
15. References
1. CRC Handbook of Chemistry and Physics. 93rd ed. CRC Press: Boca
Raton, FL, 2012; p 6-8.
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SOP No. QC-19-09
Date Revised 03-05-18
Page 6 of 8
Sample Pipette Verification Record Sheet
OPP Microbiology Laboratory
Verification Date(s): Initials:
Manufacturer
Model No.
Serial No(s).
Volume
Range
In-house Verification
Volume(s)
Verification
Status1
Corrective
Action
(Y/N)
X12523D
Gilson
M10
X12649D
1-10 nL
10 nL
BH15232
MH05165
Gilson
M100
GG05125
10-100 nL
100 nL
GG05127
AE10020
10 nL (125 nL tip)
100 nL (1250 |iL tip)
Gilson
Distriman
BD10010
1-1250 nL
U10048H
900 (12.5 mL tip)
Eppendorf
Repeater M4
H35817G
20 nL-
10 mL
30 nL (1 mL tip)
100 nL (5 mL tip)
900 ^L (10 ml tip)
10 mL (50 mL tip)
A0504243A
L0508039A
C0823986A
J0753884A
C0825980A
Rainin
L-1000
C0823596A
100-
1000 nL
H0101474A
1000 nL
H0100977A
J0902334A
J0908624A
D0303509A
E0301364A
A0510192A
L0509218A
Rainin
L-200
C0821962A
20-200 nL
100 nL
J0750805A
C0820542A
C0820661A
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SOP No. QC-19-09
Date Revised 03-05-18
Page 7 of 8
Manufacturer
Model No.
Serial No(s).
Volume
Range
In-house Verification
Volume(s)
Verification
Status1
Corrective
Action
(Y/N)
G0101809A
G0102379A
J0902612A
C0401654A
Rainin
L-100
C0825210A
10-100 nL
100 nL
C0825238A
A0507382A
C0822081A
J0724932A
C0822312A
Rainin
L-20
C0825315A
2-20 nL
10 nL
F0100492A
F0100448A
J0903005A
C0400398A
Rainin
L-2
H0100116A
0.1-2 nL
2 nL
HO100003A
Rainin
L-5000
D1080497A
0.5-5 mL
5 mL
Rainin
L-10000
A1058487A
1-10 mL
10 mL
Rainin
L-20000
L0931886A
2-20 mL
20 mL
Verification status = PASS if percent inaccuracy from gravimetric analysis is within ±5% or if vendor calibration
certificate indicates acceptable results. If verification status = FAIL, record "yes" in Corrective Action column and
fill in Action Taken below.
Corrective Actions:
Pipette Serial Number Action Taken (QAO must be notified)
1)
2. ) I
3. ) I
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SOP No. QC-19-09
Date Revised 03-05-18
Page 8 of 8
Sample Pipette Inventory
OPP Microbiology Laboratory
MLB Pipette Inv eurory
Mods' Xo
Serial Xofij
X22523D
Gilson
Ml®
X i 2 64 9D
BH15232
MKC5] 55
Gilson
Ml 00
GG3512-
GG05125
AE1 0020
Gilson
Dtsbiman
LD 0552-
U3 00-8K
Eppendarf
F.ep eater M4
K35S1~G
AO 53-243A
L050S33SA
CDS239SSA
J0~55SS4 A
C0S25SS0A
C0S23595A
Rairin
L-1000
EO'iO 14 "'4 A
HO 10 05 "7 A
JO 9 C2 3 34 A
JO SOS 524 A
D0303503A
E03013S4A
E06135 83A
Manufacturer
Modi I Xo
Serial Xoiii
AOS 10192 A
L050S21SA
C0S21552A
JO "5 OS 05 A
COS 20 54 2 A
Ratlin
L-200
C0S2056;A
G010 3S09A
G01 023"3A
JO 9 C 2612 A
C 04 0165-1A
E35!5355A
Rairan
L-100
C0S2521OA
COiS 25 238 A
AC50~3S2A
CO 522 OS!A
JO "2 45 32 A
COS 2 23 12A
Ranm
L-20
C3S25315 A
F 01C 04 29 A
F010044SA
JO 90 300^ \
C 04 0 03 S SA
R at tin
L-2
HOI 001 ISA
HO 10 000 3A
Ranin
L-5000
D10SC4"A
Ranin
L - i u I u
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